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Purine synthesis
1. Potential test question: origin to critical atoms for nucleotides
Formation of Deoxyribonucleotides
1. Deoxyribonucleotides are synthesized from their corresponding ribonucleotides by the
reduction of their C2’ position rather than by their de novo synthesis from deoxyribose-
containing precursors
2. The enzymes that catalyze the formation of deoxyribonucleoties by the reduction of the
corresponding ribonucleotides are named ribonucleotide reductases (RNRs)
a. Ribonucleotide reductase- allosteric regulation (refer to figure for
CDP,UDP,GDP,ADP)
i. The synthesis of the 4 dNTPs in the amounts required for DNA synthesis is
accomplished though feedback control
1. dATP buildup causes inhibition of all nucleotides
2. dGTP buildup inhibits all except for dATP which it stimulates
3. dTTP buildup inhibits dTTP and dCTP and stimulates dGTP
a. Also stimulated by ATP
4. dCTP stimulated by ATP
3. Genetic disease
a. Enzyme phosphoribosyl transferase defect causes Oroticacid
i. When the enzyme is blocked the PRPP also builds up
1. PRPP is a positive regulator and its buildup creates a positive feedback
yielding more of the oroticacid intermediate
Nucleotide metabolism
1. Salvage pathway
a. Most cells have an active turnover of many of their nucleic acids which, through
degradative processes results in the release of adenine, guanine, and hypoxanthine
i. These free purines are reconverted to their corresponding nucleotides
through salvage pathways
b. In mammals, purines, for the most part, are salvaged by 2 different enzymes:
i. Adenine phosphoribosyltransferase (APRT) medicates AMP formation through
the transfer of adenine to PRPP with the release of PPi
1. Adenine + PRPP AMP + PPi (reversible reaction)
ii. Hypoxanthine-guaning phosphoribosyltransferase (HGPRT) catalyzes the
analogous reaction for both hypoxanthine and guanine
1. Hypoxanthine + PRPP IMP + PPi (reversible reaction)
2. Guanine + PRPP GMP + PPi (reversible reaction)
iii. Lesch-Nyhan disease is a deficiency of this transferase leads to gout, mental
retardation, self mutilation and choreoathetosis
c. Salvage pathway is very important in the creation of momoclonal antibodies for
cancer treatment as well as other diseases
i. Myeloma cells- tumor of antibody secreting cells
1. Inject antigen into an animal and select B cells that will yield highly
specific antibody for what you want
a. These cells have a finite lifespan but you can fuse with myeloma
cells to produce immortality
i. The fusion is random and not a complete 1:1 ratio of B
cell: Myeloma cells
1. Problem with the un-fused myeloma cells or M:M
fusion because they will produce the Ab they
originally produced instead of the Ab you want
ii. HGPRP cells: after fusion you switch the cells to HAT
media
1. H- ypoxanthine to account for the decreased
synthesis of purines
2. A- methotrexate (will shut down the synthesis of
the purines and thymine so you have to add
Thymine)
3. T-hymine
Catabolism of Purines
1. All purine nucleotide catabolism pathways (which vary by organism) all lead to uric acid
a. The intermediates in these processes may instead be reused to form nucleotides via
salvage reactions
b. R1P, a product of the reaction catalyzed by PNP, is isomerized by
phosphoribomutase to the PRPP precursor R5P
2. Breakdown deficiency:
a. A deficiency in the breakdown in cysteine affects Xanthine oxidase since cysteine
transfers its S to Xanthine
i. The drug allopurinol is an xanthine oxidase inhibitor causing xanthine to be
excreted instead of uric acid
3. Genetic diseases of the breakdown pathway
a. Deaminase deficiency and deficiency in ribose removal causes an elevation in
purines and an immunodeficiency
i. The elevated adenosine feedbacks and inhibits the formation of
deoxynucleotides for DNA synthesis
1. This affects rapidly dividing tissues such as immune cells
2. Adenosine is a stress para-hormone
a. Overview
i. G proteins are molecular switches: they are active when GTP is bound to
them and inactive when GDP is bound
ii. They also have intrinsic, although weak, GTPase activity, as such, they
eventually turn themselves off when they hydrolyze the bound GTP to GDP
iii. In the active state, they can stimulate enzymes, ion channels, and affect the
cytoskeleton and vesicular trafficking
b. Mechanism of G-proteins
a. The α subunit is the GTPase, while the β γ dimer is involved with membrane
localization, protein and receptor association
i. Although GTPase acivity of α is not as slow as in the small G proteins, it still
has a GAP, called negative regulator of G protein signaling (RGS)
ii. The receptor itself acts as the exchange factor which facilitates the exchange
of GDP to GTP
i. Again the hormone binds to the receptor causing the helices to rotate
opening the cleft allowing GTP to come in and dissociate GDP, activating the
alpha subunit which can then inhibit the adenylate cyclase
*There are several different types of RasGEFs, each with its own mechanism of control. In neurons, one
Ras GEF (RafGRF1) is directly activated by Ca and a Ca binding protein called Calmodulin. On the other
hand, Phosphorylation by a cAMP-dependent protein kinase will inhibit this GNP (GEF). Therefore there
are a lot of different proteins that can feed into this system depending on the specific tissue type
*Another GEF CNrasGEF, directly binds cyclic nucleotides and is stimulated by them
*A third RasGEF, called mammalian SOS is activated indirectly: receptor or soluble TK either
Phosphorylate themselves or some docking protein .
i. However, cytosolic levels are kept extremely low, because many cellular
processes are dramatically affected by calcium
ii. As such, hormones can regulate these cellular functions by controlling the
cytoplasmic calcium concentrations
iii. The most direct mechanism for elevating calcium would be for hormones to
activate ligand-gated calcium channels, such as the glutamate receptor; by
merely opening these channels hormones would cause external calcium to
flood the cytoplasm
2. There are several PLC groups distinguished by both their structure and
their hormone regulation
a. IP3 binding opens the channel and allows the calcium to enter
the cytoplasm
b. Phospholipids
1. Glycerol backbone
3. A phosphate on the 3rd position which bridges the glycerol and some
type of head group
ii. Tyrosine Kinases both the RTK or the CR which have soluble TK that associate
with them, both of those through Tyrosine phosphorylation and binding will
activate phospholipase C gamma
1. The hormones that use G protein coupled receptors will use either Gq
to activate phospholipase C beta or the other Gprotein
c. Sphingolipids
a. Phospholipase A2
ii. Overall… TK activate Ras, Ras activates MAP kinase, MAP Kinase
phosphorylates and activates Phospholipase A2
1. For Ras signaling information refer to figure 1 of Metabolism packet
b. The 2nd fatty acid of phosphatidylinositol is arachidonic acid, which is a precursor for
the eicosanoids (example. The prostiglandins)
i. The arachidonic acid can be active and bind to various molecules and affect
their activity; therefore, it can act in 2 ways:
1. As a secondary messenger
6. While studying hormones that activated TK it was found that you activated a phosphotidyl
inositol 3 prime kinase
a. This also has a phosphor-tyrosine binding site and is activated by directly binding to
the Receptor TK
Regulation By Phosphatases
a. PKA will phosphorylate glycogen synthase which will shut it off partially because
there are multiple phosphorylation sites and each of those are additive
c. The phosphorylase kinase will then phosphorylate and activate the glycogen
phosphorylase and cause it to become fully active an no longer restricted by
allosteric regulation.
2. Effects of Calcium/Calmodulin
a. In Muscle
ii. The calcium can activate a number of protein kinases: PKC and CaMKII and
they can additionally phosphorylate the glycogen synthase to further
enhance the inhibitions.
b. In Liver
i. Vasopressin stimulates glycogen breakdown, it binds to a g protein coupled
receptor,Gq
2. IP3 has a calcium channel in the ER that will release the Calcium from
that.
3. Insulin
i. The insulin receptor can tyrosine phosphorylate the receptor for epinephrine
inhibiting it.
b. The insulin receptor can also activate a G protein, Gi which will inhibit the adenylate
cyclase.
f. Insulin can go through the MAPkinase cascade which will attract the Ras GEF(Ras
exchange factor) and the Ras will activate Raf which will activat the MapKinase.
S6KII can phosphorylate and reactivate protein phosphastase 1 which will then
remove the phosphates that PKA and these other kinases all ready put on.
a. So the fat cell pre-digests the fatty acids and break them down into Acetyl CoA.
2. It could simply hydrolyze Acetyl CoA and dump it into the blood however its vinger and it
is quite acidic and the larger the chain the less acidic the fatty acid is and to compromise
between acidity and hydrophobicity is going to be a dimer.
a. We have a thiolase that is going to couple 2 Acetyl CoA’s into Acetoacetyl-CoA and
in order to convert the energy we have a 3rd acetyl CoA come on to form Hydroxy
Methyl glutarate (HMGCoA) and then we split of the acetyl CoA to form acetoacetate
which is the first of a group of compounds refered to as ketone bodies.
b. We can reduce the carbonyl on the 3rd carbon of Aceto-acetate to a hydroxyl group,
which produces beta-hydroxybutyrate which is another ketone body.
c. You can also remove the Carboxilic acid of Acetoacetate, which will form acetone.
i. Then get Acetoacetyl-Coa which can be split by a thiolase and the 2 Acetyl
CoAs can be fed back into the TCA cycle.
3. Acetone is noticeable in diabetics who have ketoacidosis which are individuals who did not
take there insulin so that the cells can’t get the glucose it needs and the cells send out a
signal because they are starving and the body starts to breakdown Fat, and the ketone
levels will rise to very high levels.
a. So when you are fasting and you want to break down fatty acids and convert them
into ketone bodies for transport NAD levels rise and they activate SIRT and SIRT
deacytelates the HMGCoA synthase which then allows for the production of the
ketone bodies.
a. Glucagon is going to use cAMP and the genes cortisol induces are italicized because
cortisol is a steroid and is going to act by gene induction.
2. We want to break down protein so the first thing we want to do is to stop protein
synthesis.
3. Insulin, which is an anabolic hormone, is going to stimulate protein synthesis and it does it
through the mTOR and S6KI pathway.
4. Cortisol induces a gene called RED1, which will interfere with the insulin stimulation of
protein synthesis.
5. Cortisol also induces a muscle specific E3 ligase, is important in that it choses the
substrate, which is going to target the muscle filaments.
a. FOX01 is a co-activator with the cortisol receptor in inducing the muscle specific E3
ligase gene.
7. The protein is tagged it goes to the proteosome and is broken down into amino acids.
8. The first thing that happens is that the nitrogen is going to be stripped off and detoxified.
a. The cortisol will induce the arginosuccinate lyase and the arginase.
b. In addition the Carbamoyl phosphate synthase, Orn carboxyl transferase, and the
arginosuccinate lyase are all acetylated and inhibited by the acetylation.
9. So if you are fasting, you need to break down protein and you are going to have ammonia
and you need to detoxify it so the elevated NAD will activate the SIRT that will remove the
inhibitory acetate so that now these enzymes in the urea cycle will be activated.
a. The 3 steps require 4 enzymes the last step in glycolysis or the first step in
gluconeogenosis requires 2 steps.
b. The 4 enzymes: the pyruvate carboxylase, PEP carboxy kinase, F1, 6BP
phosphatase (in reverse its PFK2 which is the committed step in glycolysis)and G6P
phosphatase and all 4 of these enzymes are induced by cortisol.
11. You don’t want both glycolysis and gluconeogensis going on at the same time so
the other half is to not only inducing gluconeogensis but to shut off glycolysis and that is
the job of Glucagon.
a. Glucagon: PKA will phosphorylate the PFK2, which produced the allosteric activator
of the enzyme and pyruvate kinase, and it inhibits both of those.
c. Cortisol induces the enzymes for gluconeogensis and Glucagon uses PKA to inhibit
the enzymes of glycolysis
12. Many of the mRNA’s of the enzymes have short half-lives and are marked by AU rich
tails, which attract the AUF1, which recruits nucleases that chews up the mRNA.
13. PKA can phosphorylate the AUF1 binding protein and neutralize it so that it does not
recruit the nuclease for the gluconeogenic enzymes and we increase the half-life of the
mRNA.
14.Cortisol and Glucagon work hand in hand with each other in order to synthesis glucose.
Protein Synthesis(Diagram 2)
1. eIF-4F is responsible for binding the mRNA, getting all the kinks out of it and bringing it to
the small subunit.
b. Eif-4E, which is the cap binding protein and this, is where the control is going to be.
2. There is a 4E binding protein that will bind the cap and keep it away from the complex
thereby putting the breaks on translation.
b. The kinase that phosphorylates the 4E binding protein is known as the S6K1.
a. Rheb will bind to mTOR and activate it and mTOR will then phosphorylate and
activate S6K1 that will phosphorylate and inactivate the 4E binding protein.
b. mTOR will also phosphorylate a regulatory subunit that will inhibit PP2A so that the
phosphate will not be removed.
5. Like all G proteins Rheb is a GTPase and there is a RhebGAP that facilities the hydrolysis of
GTP thereby turning Rheb off. Rheb Gap is where everything comes in.
a. Insulin is going to activate PKB, PKB will phosphorylate Rheb Gap and inhibiting and
shutting off the GAP (Rheb Gap has both stimulatory and inhibitory phosphorylation
sites).
b. No Gap then the GTP is not hydrolyzed Rheb remains active an mTOR remains
active you get the 4E binding protein neutralized an translation proceeds which is
what you want because insulin is an anabolic hormone it is going to stimulate
protein synthesis just as it did glycogen synthesis and it stimulates lipid synthesis.
6. The growth factor is going to activate the MAP kinase also phosphorylates Rheb GAP and
inhibits it and shuts it off which then prevent Rheb GaP from inhibiting Rheb and leads to
translation.
a. MAPkinase can also activate translation indirectly by activating SDKII, which can
also phosphorylate Rheb.
7. Insulin is apposed by PKA which phosphorylates LKB1 and this is upstream of AMPK (which
is the kinase that is activated by AMP) and you don’t have excessive amount of AMP
unless energy levels are very low, and if you have low energy levels you don’t want to be
expending them by synthesizing proteins.
a. So if AMP levels are elevated then AMPK is activated and will phosphorylate Rheb
GAP on a stimulatory site enhancing its activity causing it to hydrolyze the GTP
thereby shutting Rheb off, you don’t get mTOR activation or PP2A inhibition so the
PP2A will remove the phosphate and the binding protein will sequester 4E and
translation stops.
b. Glucagon can do the same thing it activities PKA which will phosphorylate LKB1
which can phosphorylate and activate the AMP kinase.
8. In fasting you have elevation of NAD which activates a deacetylase, SIRT, and it removes
the acetate off of LKB1 and that will activate the AMP kinase.
9. ROS can activate the kinase ATM. And if you have cell stress you want to put the hold on
translation. And ATM is going to also phosphorylate LKB1 and shut down protein synthesis.
a. If you have plenty of Glucose you should have plenty of energy and Glucose 6
phosphate dehydrogenase will bind Rheb and block its interaction with mTOR.
11.Amino Acids act through another G-protein called RAG which feeds into this system.
Introduction to photosynthesis
1. Photosynthesis is divided into light and dark reactions
a. Light reactions
i. Must occur in light since it involves the absorption of photons
1. The energy from photons is used to strip H from H2O yielding O2
a. The H go to synthesize NADPH which is fed into the dark
reactions
b. In the process of doing this, the light reactions will pump H into
the chloroplast inter-membrane space
i. The H returns through an ATPase generating ATP
ii. These reactions can be further subdivided into: (refer to figure)
1. Photosystem I
2. Photosystem II (This reaction occurs before Photosystem I)
a. Absorbs a photon to split H2O
b. Dark reactions
i. Can occur in light or dark, but the materials needed (NADPH and ATP) come
from the light requiring reactions
1. These reactions reduce CO2 to carbohydrates (CH2O…)
April 1, 2011
Photosynthesis continued
LIGHT reactions
1. Energy requirements for light and dark reactions
a. Photosystem I will synthesize NADPH
i. 2NADP + 2H2O 2NADPH + O2 (438kJ/mol free energy)
ii. You get ~1ATP per 3H pumped = 2ATP per H2O molecule
1. But you start with 2 H2O meaning 4ATP produced (122kJ/mol)
a. Total is 560kJ per O2 molecule
2. Chloroplast
a. Components
i. Stroma = matrix
ii. Grana = stacked membranes
iii. Stroma lamellae = un-stacked membranes
iv. Thylakoid lumen = between membranes (equivalent to mit inter-membrane
space)
v. Photocomplex II- within the stacked membrane
vi. Photocomplex I- usually within stroma lamellae
1. Requires less energy for activation than photosystem II, this is the
reason for their separate distribution in the membrane
b. Can accept H from NADPH to make H2O
i. Only accounts for ~10% of the respiration in plants (called chlororespiration)
1. Remaining ~90% still from mitochondria
ii. Plastid terminal oxidase (PTOX) carries out chlororespiration
1. No gradient, this enzyme couples the H to O
2. Mitochondria can also use an enzyme like this called AOX
a. In mitochondria it works as a metabolic clutch
b. Plants use TCA cycle to generate intermediates in synthesis
(carbon skeletons) not for energy since it comes from light
c. AOX also functions to generate heat in fluorgenic tissue
i. Many plants are pollinated by heat
3. In chloroplast PTOX tends to occur during dehydration or in the dark
where you would not expect active photosynthesis
a. This may function to keep chloroplasts primed for
photosynthesis when it is again available
4. POTX can also dispose of excess electrons anywhere not just in the
chloroplast
5. POTX can generate NAD or NADP if you have them in excess from
other pathways
3. Light can create reactive oxygen intermediates
a. Under intense light you stimulate pq
i. Reduced pq activates a kinase to phosphorylate the light harvesting complex
to reduce the absorption of light and to initiate the shift of photosystem I next
to photosystem II
4. Absorption of light
a. Light harvesting complex II
i. Comprised of:
1. Chlorophyll A (7 of them) and chlorophyll B (5 of them)
2. 2 Lutins which act as cross braces
3. CP24 and CP29 proteins which are involved in nonphotochemical
quenching
a. Prevents over stimulation by excess light
4. ***This complex occurs in triplicates ***
a. Thus, there are 7x3=21 chlorophyll A, 15 chlorophyll B, and 6
Lutins
ii. Differences from Iron structures:
1. Chlorophyll has long chain alcohols, an extra ring, and no iron
5. Efficiency of photosynthesis (light harvesting complex):
a. Planks law
i. E = h v
1. Planks constant x frequency of wavelength
ii. E = (6.626 X 10^-34 Js) (2.998 x 10^8 m/s) (6.023 x 10^23) / (680 x 10^-
9m)
iii. E = 126 kJ / Einstein x 8 photons
iv. E = 1408 kJ
1. From this you can capture 560 (~40%)
a. Actual efficiency is ~25%
6. Refer to figure 22-13 strayer biochemistry 3rd ed (non-cyclic pathway
7. Refer to figure for violaxanthin
a. pH indicator system used to protect against photo oxidative damage
8. Refer to figure Cyclic electron flow pathway
a. Bypasses photosystem II thus no O2 or NADPH produced
i. You pump H making a large amount of ATP (10ATP instead of the usual 4ATP)
b. Used when there is insufficient NADP or you need more ATP
DARK Reactions (Calvin cycle)
1. Dark reactions use the NADPH and ATP generated in the light reactions
2. Cycle consists of 3 major processes
a. Carboxylation of Ribulose-1,5-bisphosphate with CO2 and H2O two 3-
phosphoglycerate
b. Reduction of two 3-phosphoglycerates with 2ATP and 2NADPH triose phosphate
i. Trios phosphate splits off some to generate sucrose and starch while the rest
is regenerated
c. Regeneration of trios phosphate with ATP ribulose-1,5-bisphosphate
3. Refer to circle figure with ribulose-5-phosphate at the top
April 4, 2011
Dark Reactions, Calvin Cycle Continued
1. Regulation
a. Step 1: Ribulose-1,5-bisphosphate + CO2 2 3-PG
i. Done by the ribulose bisphosphate carboxylase (rubisco)
1. Rubisco comprises ~30% of all plant protein
2. Rubisco consists of 16 subunits (8- 56kD subunits and 8- 14kD
subunits)
3. Free energy of the reaction is -35kJ per mole
4. Has a very high affinity for CO2 but a very low catalytic efficiency (3CO2
per second)
5. Heavily regulated by various factors:
a. pH- Rubisco makes sure you do not fix CO2 unless you have light
present
b. Mg2+ - If H is pumped out of the stoma it is replaced by Mg2+ to
maintain the charge
i. You only have the Mg2+ when active photosynthesis is
taking place
c. Allosterically regulated by a small sugar (CA1P) which acts as a
transition state analog only synthesized in the dark where
photosynthesis is not occurring
d. The protein Rubisco activase is a catalytic chaperone that can
counteract the activity of CA1P
i. Helps the enzyme achieve optimum activity without
photosynthesis
ii. Rubisco activase is regulated by ADP:ATP
1. When the ratio is 1:1 (typical of the dark) it is
inactive
2. When the ratio is 1:2-3 (typical of the light) it is
activated
b. Phosphatase enzyme regulation
i. Regulated by disulfide bonds
1. With the disulfide bond they are inactive
2. You couple enzymes in the Calvin cycle to the light reactions
2. Efficiency
a. 6CO2 + 18ATP + 12(NADPH + H+) glucose + 18ADP + 12 NADP
i. Energy consumed: 18 ATP = 30.5 kJ per mole per ATP = 549kJ/mol
1. 12 NADPH x 219kJ each =2628
2. Total = 3177 per mole of glucose used
3. If you burn that glucose CO2 + H2O = 2823kJ/mol
3. Other ways to fix Carbon:
a. Bacteria
i. Acetogenic bacteria
1. Take 2 CO2 and progressively reduce them to acetyl CoA and H2O
ii. 3-Hydroxypropinate cycle
1. Start with 2 Acetyl CoA + 2HCO3 Malonyl CoA
2. Reduce Malonyl CoA to a hydroxyl group 3-hydroxypropinate
3. Couple 3-hydroxypropinate to CoA 2 Propionyl CoA
a. 1 Propionyl CoA is converted to Succinyl CoA for the TCA cycle
i. Succinyl CoA Malate coupled to CoA Regenerating
Acetyl CoA and Glyoxylate as a byproduct
b. The Glyoxylate byproduct is attached to the 2nd Propionyl CoA
i. Rearrange to form Citramalyl-CoA
ii. Citramalyl CoA is split into pyruvate and Acetyl CoA
4. Rubisco and its oxidizing capability (refer to figure 3 of photosynthesis packet)
a. Photorespiration- occurs when the stomata are closed
i. When closed the plant switches to oxidation depleting the cell of oxygen and
increasing the amount of CO2
ii. The limiting factor in plant growth
1. Protects against photooxidative damage when excess O2 is present
2. Essential for getting N from the environment
5. Integration of Calvin cycle and synthesis of Fatty acids (refer to figure 7 of photosynthesis
packet)
a. Calvin cycle recycles CO2
i. By linking Calvin cycle with glycolysis the same 5 hexose phosphates will give
you 6 pentose phosphates, 12 acetyl CoA and 6CO2
6. Heat and the stoma
a. Must close during high heat to keep water but this also limits the intake of CO2
i. So all of the ATP and NADP from the light reactions back up since there is no
CO2 to produce
b. The C4 plants can store the CO2 in 4 carbon molecules to prevent this backup (refer
to figure 4 in photosynthesis packet)
i. Mesophyll cells are next to the stomata and have access to the CO2
ii. Bundle sheath cells are more internal
1. There are connections between the 2 called plasmodesmata
a. Called Kranz anatomy
i. Variations in Kranz anatomy
1. Some cells do this process in a single elongated
cell, instead of in mesophyll and bundle sheath
cells
iii. CO2 is stored in the cell as malate (in some cases, but it can also be stored as
other substances such as aspartic acid)
1. When the stroma close and the CO2 begins to fall it is transferred to
the bundle sheath cells to keep the Calvin cycle going and produces
pyruvate
iv. CO2 cannot typically be stored as bicarbonate because it alters the pH
1. This takes place in some algae
7. Rubisco can use either CO2 or O2
a. During the evolution CO2 levels have fallen while O2 levels have risen
i. The O2 can affectively compete with the CO2
1. So the cells increase the local CO2 concentration to steer the reaction
toward CO2
April 6, 2011
C4 Plants Continued
1. Review/Continuation from last class: (Refer to figure 4 of photosynthesis packet)
a. Enzyme regulation:
i. PEP carboxylase and the Malate dehydrogenase are two key regulation
locations
2. Pyruvate Dikinase is the rate limiting step and is regulated by a dedicated PPDK (Refer to
figure 5 of photosynthesis packet)
a. PPDK can act as a phosphatase
i. 2 phosphorylation sites:
1. Histidine site functions to transfer the phosphate to the pyruvate to
form phosphoenolpyruvate
2. Threonine site phosphorylation makes the enzyme inactive
b. PPDK regulatory protein can also acts as a kinase in the dark
i. The TAP will be consumed in the absence of light to form ADP which PPDK will
use
3. PEP CO2ase
a. Allosterically inhibited by malate
b. PEP CO2ase-PO4
i. The kinase that catalyzes this PO4 addition is activated by light